Madridge Journal of Internal and Emergency Medicine

ISSN: 2638-1621

International Translational and Regenerative Medicine Conference

April 25-27, 2018, Rome, Italy
Scientific Session Abstracts
DOI: 10.18689/2638-1621.a1.002

Inhibition of Breast Cancer Bone Metastasis and Pancreatic and Colon Cancer by Synthetic Curcumin Analogs

Mamoru Shoji

Emory University School of Medicine and Winship Cancer Institute, USA

Curcumin (diferuloylmethane) is a β-diketone constituent of the turmeric. It is used as a spice to give a specific flavor and yellow color to curry. However, its clinical efficacy is poor because of itʼs low solubility. He worked with professors Liotta and Snyder at the Chemistry Department to synthesize a series of novel monocarbonyl analogs of curcumin (MACs) approximately 100 analogs including EF24, EF31 and UBS109. Dr. Shojiʼs laboratory and the NCI tested the analogs for the anticancer activity.

The NCI determined the mean growth inhibitory concentration (GI-50) of EF24, curcumin and cisplatin on the NCI-60 cancer cell panel, which are 0.7 µM, 7.3 µM and 9.5 µM, respectively. MACs do not kill normal breast cells MCF-10A but kill all cancer cells tested (KB-3-1, TU212, MiaPaCa, SE-Mel-28, RPMI-7951, and MDA-MB-231 cells) at concentrations (0-20 µM) [Zhu S, Current Cancer Drug Targets, 2014]. MACs inhibit NF-κB by inhibiting IKK-α and IKK-β.

UBS109 inhibited breast cancer metastasis and osteolysis by inhibiting osteoclasts precursors and osteoclasts, but promotes new bone formation by stimulating osteoblast activation. UBS109 and EF24 inhibited four pancreatic cancer cell lines 100% at less than 1.25 µM, whereas gemcitabine did not up to 20 µM. UBS109 significantly inhibited MiaPaCa-2 pancreatic cancer xenografts and colon cancer (HT-29 and HCT-116) xenografts in mice at 25 mg/kg, iv once a week better than a combination of oxaliplatin (5 mg/kg) and 5FU (30 mg/kg) iv.

Dr. Mamoru Shoji obtained his Medical Degree from the Hokkaido University, Japan, and completed internships at the US Naval Hospital, Yokosuka, Japan and the University of Pennsylvania in Philadelphia, residency in internal medicine at the Lahey Clinic, Boston, fellowship training in immunology at the Peter Bent Brigham and Robert Breck Brigham Hospitals (mentor, John R. David, MD), Harvard Medical School in Boston, in tumor immunology at the University of Minnesota (mentor, Charles F. Mckhann, MD from Massachusetts General Hospital) in Minneapolis, followed by fellowship in Hematology and Medical Oncology at Emory University (mentor, Charles M. Huguley, Jr, MD).

Immune Modulating Properties of the Anti-Cancer Preparation NSC631570

Wassil Nowicky

Nowicky Pharma and Ukrainian Anti-Cancer Institute, Austria

Unusual for an anticancer agent NSC-631570 possesses some distinct immune properties [35, 49]. It was Prof. AndrejsLiepins of the St. Johnʼs Memorial University, St. Johnʼs, Canada who first pointed to this interesting fact. In the work with the C57BL/6 mice he revealed NSC-631570 to be an effective biological response modifier (BRM). After incubation with NSC-631570 the lytic activity of the splenic lymphocytes from the alloimmunised mice increased up to 48 fold (fig. 4).

The immune modulating effect of NSC-631570 was studied in several studies in mice. Repeated subcutaneous injections of NSC-631570 to mice infected with the twofold LD50 of E. coli, S. aureus, or influenza virus increased the survival rate of the animals significantly. When human lymphocytes were incubated with phytohemagglutinin (PHA) and NSC-631570, increased absorption of 3H-thymidin in the cells was observed. The authors point out the strong synergetic effect of NSC-631570 and phytohemagglutinin. NCS-631570 induces immunogenic death of B16 melanoma cells and could restore antitumor activity of hypoxia-polarized macrophages. It suggests that NSC-631570 can be used for multimodal tumor therapy not only to kill the tumor cells, but also to stimulate a specific immune response to keep residual tumor (stem) cells and metastases under control. To investigate the adjuvant and immunomodulatory effects of the S.aureus cytoplasmic membrane extraction (CPM) in mono- and combined anticancer therapy outbred mice were transplanted with ascite and solid form of Ehrlich carcinoma followed by six day course of bacterial polymer used alone or in combination with cytotoxic anticancer drug NSC-631570. To estimate adjuvant effect tumor growth dynamics were evaluated, to characterize immunomodulating effect the number of circulating mononuclear phagocytes and their hagocytic activity were analyzed by flow cytometry. Our results suggest synergistic effect of PAMP and antineoplastic drug NSC-631570, that was accompanied by positive immunomodulation. The effects of cancer-selective drug NSC-631570 (Ukrain) used alone and in combination with pathogen-associated polymers of Gram-positive (peptidoglycan, lipoteichoic acid, and cytoplasmic membrane extraction of Staphylococcus aureus) and Gram-negative (Escherichia coli lipopolysaccharide) bacteria on mouse peritoneal macrophage metabolic activity in vitro are investigated. It is shown that NSC-631570, as used alone, causes a moderate enchancement of oxidative metabolism and arginase activity of intact peritoneal macrophages. The co-modulatory effect of the preparation depends on the initial functional state of phagocytes.

Dr. Wassil Nowicky Diplom-Ingenieur, Doctor scientiae technicorum, DDDr. h. c., Director of “Nowicky Pharma” and President of the Ukrainian Anti-Cancer Institute (Vienna, Austria). He has finished his study at the Radio technical Faculty of the Technical University of Lviv (Ukraine) with the end of 1955 with graduation to “Diplomingeniueur” in 1960 which title was nostrificated in Austria in 1975.
Dr. Wassil became the very first scientist in the development of the anticancer protonic therapy and is the inventor of the preparation against cancer with a selective effect on basis of celandine alkaloids “NSC-631570”. He used the factor that cancer cells are more negative charged than normal cells and invented the Celandine alkaloid with a positive charge thanks to which it accumulates in cancer cells very fast.
Thus, Dr. Nowicky is invited as an Honorable Speaker to take part in many scientific international congresses and conferences in USA, Australia, Japan, UAE, and Europe. Author of over 300 scientific articles dedicated to cancer research.
Dr. Wassil Nowicky is a real member of the New York Academy of Sciences, member of the European Union for applied immunology and of the American Association for scientific progress, honorary doctor of the JankaKupala University in Hrodno, doctor “honoris causa” of the Open international university on complex medicine in Colombo, honorary member of the Austrian Society of a name od Albert Schweizer. He has received the award for merits of National guild of pharmasists of America and the award of Austrian Society of sanitary, hygiene and public health services and others.

Synthetic Aromatse Inhibitors in the Treatment of Estrogen Dependent Breast Cancer

Shagufta Waseem* and Irshad Ahmad

Mathematics and Natural Sciences, School of Arts and Sciences, American University of Ras Al Khaimah, UAE

Breast cancer is the most common form of cancer present in women worldwide and is the second leading cause of death after lung cancer.1 In estrogen-dependent breast cancer, estrogen plays a significant role in the stimulation of breast cancer cell proliferation.2 Two main approaches have been developed to control or block the pathological activity of estrogens. The first approach involves the design and synthesis of estrogen receptor antagonist and inhibition of enzyme aromatase is the second approach for the development of new agents for the breast cancer treatment.3-5 Aromatase, an enzyme complex present in breast tissues, plays a substantial role in the biosynthesis of important endogenous estrogens from androgens. The source of oestrogen production in breast cancer tissues is intra-tumoral aromatase, and inhibition of aromatase may inhibit the growth stimulating effect of estrogens in breast cancer tissues. Consequently, aromatase is considered a useful therapeutic target in the treatment and prevention of estrogen-dependent breast cancer. Recently, different natural products and synthetic compounds have been rapidly developed, studied, and evaluated for aromatase inhibitory activity. Aromatase inhibitors are classified into two categories on the basis of their chemical structures, i.e., Steroidal and nonsteroidal aromatase inhibitors.6 This presentation will emphasize the potent role of synthetic steroidal and nonsteroidal aromatase inhibitors for the treatment of breast cancer.

Dr. Shagufta joined the American University of Ras Al Khaimah as an Assistant Professor of Chemistry in the School of Arts and Sciences in August 2014. Before joining AURAK, she worked as an Adjunct Assistant Professor of Chemistry at the University of Modern Sciences, Dubai and American University of Ras Al Khaimah, UAE.
She worked as a Postdoctoral Researcher Associate at the Department of Chemistry and Biochemistry, Oklahoma University, USA. She developed the noble drug delivery system for breast cancer drugs using carbon nanotubes and acquired the significant experience in nanotechnology and synthetic organic chemistry. She was appointed as a Postdoctoral Research Fellow and Visiting Scientist at Leiden/Amsterdam Centre for Drug Research (LACDR), Leiden, and the Netherlands. Her research interest was in silico prediction and clinical evaluation of the cardiotoxicity of drug candidates. She was focused to identify chemical substructures as ‘chemical alertsʼ that interact with this hERG channel.
She received a Ph.D. under the prestigious CSIR-JRF and SRF research fellowship in Chemistry from Central Drug Research Institute (CDRI)/Lucknow University, India in 2008, her PhD research work was in the field of estrogens and antiestrogens, design and synthesis of steroidal and non-steroidal tissue selective estrogen receptor modulators (SERMs) for breast cancer, 3D-QSAR CoMFA and CoMSIA studies and analysis of pharmaceutical important molecules. She has published 20 articles in peer-reviewed International journals of Royal Society of Chemistry, Elsevier, Wiley and Springer. And she teaches courses such as General chemistry, Organic Chemistry, Chemistry in Everyday Life, and Spectroscopy along with laboratory courses.

Inhibition of NF-κB Pathway Attenuates the Multiple Organ Dysfunction Associated with Polymicrobial Sepsis in Mice with Pre-Existing Type 2 Diabetes Mellitus (T2DM)

Sura Yahia Yosef Al Zoub1, J Chen1, L Martin1, C Murphy1, C Thiemermann1, F Chiazza2, D Collotta2 and M Collino2

1Centre for Translational medicine and Therapeutics, Queen Mary University of London, UK
2Department of Drug Science and Technology, University of Turin, Italy

Introduction: Patients with diabetes are at higher risk of infections and sepsis. Activation of nuclear factor-κB (NF-κB) plays a substantial role in the pathophysiology of sepsis and diabetes. Here we investigate i) the effect of pre-existing type 2 diabetes mellitus on organ dysfunction (MOD) associated with sepsis and ii) whether inhibition of NF-κB using IKK-16 or linagliptin attenuates MOD in mice with sepsis and diabetes.

Methods: Ten-week old male C57BL/6 mice received high fat (HFD) or chow diet for 12 weeks, and were subjected to caecal ligation and puncture (CLP) or sham surgery for 24 hours. At 1 hour after CLP, mice received IKK-16, linagliptin, or vehicle.

Results: Administration of HFD resulted in a significant (i) impairment in glucose tolerance, (ii) (small) reduction in ejection fraction and, (iii) increase in alanine aminotransferase. HFD caused (Immunoplot analysis; heart) significant NF-κB pathway activation and expression of inducible nitric oxide synthase (iNOS). Mice on HFD subjected to CLP showed further (i) decline in EF, (ii) increase in serum ALT, and (iii) a significant renal dysfunction. Treatment of HFD-CLP mice withIKK-16 or linagliptin resulted in significant reduction of the CLP-induced i) MOD (cardiac/liver/renal), ii) NF-κB pathway activation, and iii) iNOS expression when compared to mice treated with vehicle.

Conclusion: Our results show that HFD results in inflammation, cardiac dysfunction, and liver injury. Moreover, a pre-existing diabetic phenotype worsened the organ injury/dysfunction associated with CLP-sepsis. Most notably, inhibition of NF-κB reduced the organ injury/dysfunction caused by sepsis in animals with pre-existing T2DM.

Dr. Sura Al Zoubi is a clinical pharmacist who is doing a PhD in pharmacology at the department of the transnational medicine and therapeutics, the William Harvey Research Institute (WHRI), Queen Mary University of London, UK. Her main research interest is the cardiac dysfunction associated with sepsis and the effect of type 2 diabetes mellitus on the pathophysiology of cardiac dysfunction in sepsis. She has been working on the effect of NF-κB pathway inhibition on the multiple organ dysfunctions in sepsis. She has presented her work in many national and international conferences.

Therapeutic Potential of Quinazoline Derivatives as Anticancer Agents

Irshad Ahmad* and Shagufta

Mathematics and Natural Sciences, School of Arts and Sciences, American University of Ras Al Khaimah, UAE

Cancer is one of the major causes of worldwide human mortality. It is estimated that about 1 688 780 new cancer cases will be diagnosed in 2017. 1A wide range of cytotoxic drugs are available on the market, and several compounds are in different phases of clinical trials. 2Many studies suggest that these cytotoxic molecules are also associated with different types of adverse side effects; therefore researchers around the globe are involved in the development of more efficient and safer anticancer drugs. The heterocycles are widely investigated bioactive molecules and are considered important synthetic targets for the development of novel therapeutic agents. 3In recent years, quinazoline and its derivatives have been considered as a novel class of cancer chemotherapeutic agents that show promising activity against different tumors.4This presentation will comprehensively highlight the recent developments concerning the anticancer activity of quinazoline derivatives as well as offer perspectives on the development of novel quinazoline derivatives as anticancer agents in the near future.

Dr. Irshad Ahmad joined the American University of Ras Al Khaimah in spring 2011 as an Assistant Professor of Chemistry. He received the masterʼs degree in chemistry from Jiwaji University in 1999. Subsequently acquired significant pharmaceutical industrial experience and developed cardio-selective beta-blocker drug molecule. He joined Central Salt and Marine Chemical Research Institute and Bhavnagar University under the sponsored project of DST and CSIR as a senior research fellow and received his PhD degree in chemistry in 2006. Subsequently, he accepted an invited scientist position in Korea Research Institute of Chemical Technology, South Korea and contributed his expertise in the field of Nanotechnology. And is a recipient of prestigious European fellowships (NWO-Rubicon & FCT) and he joined Vanʼt Hoff Institute for Molecular Sciences, University of Amsterdam, The Netherlands as a NWO Rubicon fellow (Netherlands Organization for Scientific Research, the Dutch Science Foundation), he acquired expertise in the field of supramolecular chemistry.
Afterward, he moved to the Leibniz Institute for Surface Modification, Leipzig, Germany under the Deutsche Forschungsgemeinschaft Grant, he developed “Novel ultra-fast metathesis catalyst” for the production of high quality alternating copolymers. Subsequently he joined Department of Chemistry and Biochemistry, Stephenson Life Science Research Center, University of Oklahoma, USA as a postdoctoral research associate. He developed strategies for the novel environmentally friendly reactions for the production of value added chemicals from biomass.
He is specialized in the area of chemistry, bridging the traditional disciplines of inorganic, organic and bio-organic chemistry. He contributed US and European patent for green and clean technology development. He has published peer-reviewed international research articles in the American Chemical Society (ACS), Royal Society of Chemistry (RSC) Cambridge, Elsevier Science, Wiley, and Springer journals. He has presented his research at several scientific conferences worldwide and received awards.

Upstream Open Reading Frames in Rare Human Diseases and Related Therapeutics

Ioanna A Armata

Department of Chemistry & Biochemistry and Vitalite Health Network, University of Moncton, Georges L-Dumont University Hospital Center, Canada

Translation generally initiates at the AUG codon (cAUG) marking the beginning of the coding region of a gene. Approximately 40% of human transcripts can also initiate translation within the 5ʼ untranslated region, at upstream AUG codons (uAUGs) which precede the cAUG. The presence of a uAUG within a transcript marks the beginning of an upstream open reading frame (uORF), which represents a potentially translatable nucleotide sequence. Single nucleotide polymorphisms that disrupt uAUG/uORF-mediated translation are associated with approximately 30 untreatable human diseases, such as melanoma, β-thalassemia, campomelic dysplasia and Dopa-Responsive Dystonia (DRD). Pathogenic uAUGs/uORF scan deregulate translation through two distinct molecular mechanisms: i) by reducing translation efficiency of normal protein products; and ii) by encoding mutant proteins that can trigger cellular death. Utilizing a uAUG/uORF associated with DRD, a high throughput screening assay was developed for identifying compounds that prompt the translational machinery to “bypass” pathogenic uAUGs. Screening of a small collection of bioactive compounds reveals that Salubrinal (SAL), - (a potent and selective inhibitor of eukaryotic initiation factor 2αdephosphorylation)-, promotes overriding of pathogenic uAUGs associated with DRD, as well as with melanoma predisposition, and the Van der Woude syndrome. Evaluation of SAL against uAUGs associated with additional human diseases is required to determine if SAL is a promising therapeutic agent for the group of uAUG-associated diseases. Such therapeutic agents would satisfy an unmet medical need.

Dr. Ioanna A Armata is a molecular and cellular neuroscientist, who specializes in dystonia, a heterogenic group of debilitating neurological disorders, affecting both children and adults. Her training and expertise is in the fields of gene expression, transcription factors, functional non-coding mutations, assay development for drug high throughput screenings, and the CRISPR/Cas9 system. She obtained her Ph.D. in Neuroscience at Mount Sinai School of Medicine (2008), followed by a postdoctoral fellowship (2008-2012) at Massachusetts General Hospital and Harvard Medical School, and followed by a Research Faculty I appointment at Florida State University (2013-2016). She is currently a senior Research Fellow at the University of Moncton (Canada) studying translational therapeutics for dystonia syndromes and myotonic dystrophy 1. She has recently got an affiliation with the hospital Dr. Georges-L.-Dumont University Hospital Center, Canada.

Microphysiological Systems: Organs on Chips for Drug Efficacy and Toxicity Studies

Danilo A Tagle

National Center for Advancing Translational Sciences, National Institutes of Health, USA

The current drug development process is vulnerable to poor prediction of human physiological responses and failure to predict safety and efficacy of candidate drugs using current methods accounts for a much as 90% of the attrition rate. To address this challenge in drug development, the NCATS Tissue Chip for Drug Screening program is developing alternative approaches for more reliable readouts of toxicity and efficacy. Tissue chips are bioengineered microphysiological systems utilizing chip technology and microfluidics that mimic tissue cytoarchitecture and functional units of human organs. These microfabricated devices are useful for modeling human diseases, and for studies in precision medicine and environment exposures. Tissue chips are poised to deliver a paradigm shift in drug discovery. By emulating human physiology, these chips have the potential to increase the predictive power of preclinical modeling, which in turn will move the pharmaceutical industry closer to its aspiration of clinically relevant and ultimately animal-free drug discovery. Near term uptake of these human-relevant platforms will fill gaps in current capabilities for assessing important properties of disposition, efficacy and safety liabilities. Tissue chips as novel preclinical modeling platforms offer a number of unique opportunities, with improved clinical predictions of human response being the most apparent and the greatest contribution. They may also provide a more efficient approach to mechanistic investigation, early safety liability screening and even more translationally relevant modeling of drug distribution and metabolism.

Dr. Dan Tagle is associate director for special initiatives at NCATS. He also recently served as acting director of the NCATS Office of Grants Management and Scientific Review and currently serves as executive secretary to the NCATS Advisory Council and Cures Acceleration Network Review Board. Prior to joining NCATS, Tagle was a program director for neurogenetics at the National Institute of Neurological Disorders and Stroke (NINDS), where he was involved in developing programs concerning genomics-based approaches for basic and translational research in inherited brain disorders.
Prior to joining NINDS in 2001, Tagle was an investigator and section head of molecular neurogenetics at the National Human Genome Research Institute and has been involved in the highly collaborative effort toward the positional cloning of genes for Huntingtonʼs disease, ataxia-telangiectasia and Niemann-Pick disease type C. He has served on numerous committees and advisory boards, including the editorial boards of the journals Gene and the International Journal of Biotechnology.
Tagle obtained his Ph.D. in molecular biology and genetics from Wayne State University School of Medicine in 1990. He was an NIH National Research Service Award postdoctoral fellow in human genetics in the laboratory of Francis S. Collins, M.D., Ph.D., at the University of Michigan. Tagle has authored more than 150 scientific publications and has garnered numerous awards and patents.

Meninges are a Reservoir for New Functional Neurons of the Cortex

Francesco Bifari

Laboratory of Cell Metabolism and Regenerative Medicine, Department of Medical Biotechnology and Translational Medicine, University of Milan, Italy

Whether new neurons are added in the postnatal cerebral cortex is still debated. Here, we report that the meninges of perinatal mice contain a population of neurogenic progenitors that migrate to the caudal cortex and differentiate into Satb2+ neurons in cortical layers II-IV. These neurogenic meningeal cells are generated during embryonic development between E13.5 and E16.5. The embryonically derived meningeal progenitors remain largely quiescent, and in the first days after birth, they migrate to the cortex and differentiate to cortical neurons, without further proliferation. The resulting neurons are electrically functional and integrated into local microcircuits. Using multiple lineage tracing approaches, we found that most of the meninges-derived neurons belonged to the PDGFRß-lineage, while only a small fraction of cortical cells originating from meninges was derived from GLAST+ and Nestin+ lineages generated at E13.5. Since PDGFRß is expressed by both pericytes and a subset of radial glia (RG) cells, we hypothesized that the neurogenic meningeal cells belonged to one of these cell types. Single cell transcriptomic analysis identified a PDGFRß+ meningeal cell population with a RG-like gene expression signature. We found meningeal cells with distinct transcriptome signatures characteristic of (i) neurogenic radial glia-like cells (resembling neural stem cells in the SVZ), (ii) neuronal cells, and (iii) a cell type with an intermediate phenotype, possibly representing radial glia-like meningeal cells differentiating to neuronal cells. Thus, we have identified a pool of embryonically derived radial glia-like cells present in the meninges that migrate and differentiate into functional neurons in the neonatal cerebral cortex. These findings broaden the concept of brain plasticity since they indicate that quiescent embryonically-born neural progenitors may contribute to add new functional neurons to the postnatal cortex

Dr. Francesco Bifari, MD, PhD. is Principal Investigator and Assistant Professor at the Department of Medical Biotechnology and Translational Medicine, University of Milan, Italy. (Laboratory of Cell Metabolism and Regenerative Medicine.) In 2013 he was granted by FP7-PEOPLE-2013-IEF-No626829 grant and worked at the Laboratory of Angiogenesis and Neurovascular, VIB, Leuven, Belgium (Prof. Peter Carmeliet). He is the author of 33research papers (10 first, 3 last author) and 1 book chapter. Dr. Bifari have produced several top journal publications (Cell, Cell Stem Cell, Cell Metabolism, Cancer Cell, EMBO journal) He also has a score of 1350 citations; average citations per item=45, 3; mean Impact Factor=8, 6.

Isolated Mitochondrial-Transfer into Cells: Mechanism(s) and Therapeutic Potential

Lorberboum-Galski Haya1, Kesner Eyal1, Shohami Esther2, and Rosenmann Hanna3

1Department of Biochemistry and Molecular Biology, Institute for Medical Research Israel-Canada (IMRIC), Hebrew University of Jerusalem, Israel
2Departmentof Pharmacology, Hebrew University, Israel
3The Agnes Ginges Center for Human Neurogenetics, Hadassah Hebrew University Medical Center, Israel

Mitochondrial transfer of isolated mitochondria into host cells was reported over 30 years ago, and has been well established since then. However, while mitochondrial transferʼs beneficial effects are clear, the process itself is still vague. Our attempts to characterize mitochondrial transfer into cells showed that isolated human mitochondria can be transferred into many cell types and that mitochondrial transfer to mitochondria-defected cells results in beneficial outcomes for the cells. The exogenous transformed mitochondria co-localize with endogenous mitochondria. Heparan sulfated polysaccharide molecules are crucial for this process. In addition, the outer membrane and outer membrane proteins integrity are essential for the process. Pharmacological inhibition of macropinocytosis, but not of clathrin-mediated endocytosis, impeded the transfer. Transmission electron microscopy analysis of mitochondrial transfer revealed that the isolated mitochondria interact directly with cells, which engulf the mitochondria with cellular extensions. This suggests the involvement of macropinocytosis or macropinocytosis-like mechanisms in mitochondrial transfer, in line with our pharmacological inhibition tests. The simplicity by which mitochondria can enter cells in vitro, led us to evaluate mitochondrial transfer in vivo, in a mouse model of close head injury (CHI), a type of traumatic brain injury following both motoric and cognitive abilities, as well as in an Amyloid beta (Aβ) ICV injected mouse model for Alzheimerʼs disease. In both in vivo models, mitochondrial transfer was found to have a therapeutic effect.

Dr. Haya Lorberboum-Galski is a Full Professor at the Department of Biochemistry and Molecular Biology, Faculty of Medicine, Hebrew University of Jerusalem. After receiving her Ph.D. in Biochemistry from the Hebrew University, she became a postdoctoral fellow at the Laboratory of Molecular Biology, NCI, and NIH, USA. Her main research area is Developing reagents for Targeted Human Therapy with the lately focus of mitochondrial genetic diseases. She published over 65 publications in peer-reviewed journals, review articles, edited a book on chimeric proteins and holds several patents. She served as the Head of the Department, Head of the Program for Biochemistry at the Faculty and many other committees. For the last five years she is the Chairman of the Institute for Medical Research (IMRIC) at the Faculty of Medicine, Hebrew University.

Fingolimod (FTY720) as an Anti-Multiple Sclerosis Oral Ultimate Therapy

Sara T Alrashood1, Fadilah S Aleanizy2, FulwahAlqahtani2, AhadAbushal3, Noor AlSalama3 and Ghada S Hassan4

1Department of Pharmaceutical Chemistry, King Saud University, Saudi Arabia
2Department of Pharmaceutics, King Saud University, Saudi Arabia
3College of Pharmacy, King Saud University, Saudi Arabia
4Department of Medicinal Chemistry, Mansoura University, Egypt

Multiple Sclerosis (MS) is a chronic progressive autoimmune disease. It has complex symptoms and challenges. Patients suffer the most from the inconvenience and intolerability till the old injectable disease modifying therapies. Therefore our research is highlighting the scoop on the newly available treatment using the oral therapies proceeding from their chemical nature and structure to prove their superiority to the traditional ones in controlling MS progression. This can lead to an improvement inlife quality for MS patients with the enhanced tolerability for oral drugs that is best achieved by using Fingolimod that is considered as the first approved oral drug for this disease.

FasL Pleiotropic Role on Bone Marrow Stromal Cells and its Potential Regulation by Micro RNAs

Maria Rita Rippo1, Giuliani Angelica1, Emanuela Mensà1, Fabiola Olivieri1, Antonio Domenico Procopio1 and Francesco Prattichizzo2

1Department of Clinical and Molecular Sciences, Marche Polytechnic University, Italy
2IRCCS Multimedica, Italy

Several lines of evidence suggest a pleiotropic role of FasL/CD95 on BM-MSC function. Although FasL was initially described as a T-cell-associated protein capable of inducing apoptosis by binding to its receptor Fas, a pleiotropic role in other cell populations has also been described. Fas engagement in resting T lymphocytes transduces inhibitory or costimulatory signals in a FasL dose-dependent manner, and in hematopoietic progenitors FasL receptor transduces dual apoptotic and trophic signals. Fas and FasL are expressed in freshly isolated BM–MSCs. However, cell death induction does not seem to be the Fas/FasL systemʼs main role in bone homeostasis. Fetal BM have functional extrinsic apoptotic pathways, whereas adult BM–MSCs are resistant to Fas-mediated apoptosis. FasL has a limited role in osteoblast and osteoclast apoptosis, but inhibits osteoblast differentiation in mice. We investigated the effect of FasL on BM–MSC apoptosis, proliferation, and differentiation into adipocytes to clarify the role of the Fas/FasL system in BM–MSC biology. FasL exerts a pleiotropic action on BM–MSCs depending on its concentration: low doses induce proliferation, whereas higher doses have a slight but significant apoptotic effect and, more importantly, inhibit adipogenesis; all such effects are exerted without affecting BM–MSC stemness, irrespective of dosage. Our findings show a FasL-dependent regulation of BM– MSC biology and adipogenesis, and suggest a role for FasL in conditions involving altered BM adipogenesis, such as osteoporosis in the elderly. Furthermore, our unpublished data show a modulation of several miRNAs during BM-MSC adipogenesis, Several of those regulating Fas/FasL expression, are cellular and circulating markers of aging and inflamm-aging (miR-21, miR-146a, miR-98 and miR-181a), Interestingly, plasma soluble Fas ligand concentration decrease whereas miR-21 miR-146a increase in elderly humans. Altogether these data suggest a functional axis involving miRNAs, Fas/Fasl system, bone marrow adipose tissue and aging.

Dr. Maria Rita Rippo was a student at the Department of Experimental Medicine, Policlinico Umberto I, of the University of Rome “La Sapienza 1993–1994. She obtained her Degree in Biological Sciences from the University of Rome “La Sapienza”, in 1995. She has done her PhD from 1994-2001 Department of Experimental Medicine and Biochemical Sciences, Laboratory of Signal Transduction, including a 4 months collaboration and training period at the laboratory of “Apoptose, Cancer et Immunologie”, CNRS, Villejuif, Paris, France, directed by Guido Kroemer M.D. for Immunological Sciences & In 2000 she got the Fondazione Adriano BuzzatiTraverso post-doctoral fellowship, Department of Experimental Medicine and Biochemical Sciences, University of Rome “Tor Vergata” Laboratory of Signal Transduction. She was research fellow from 2001-2002 at Polytechnic University of Marche. Worked as a Researcher and consultant (2009- 2010) at the Center of Clinical Pathology and Innovative Therapies, Italian National Research Center on Aging (INRCA-IRCCS). Obtained her Specialization in Clinical Pathology (2015) University G.dʼAnnunzio, Chieti, 70/70 cum laude. Since 2002-2016 she is a Researcher in Experimental Medicine, Pathophysiology and Clinical Pathology Department of Molecular Pathology and Innovative Therapies, Polytechnic University of Marche. And is presently an Associate Professor in Applied Medical Technologies, Department of Molecular Pathology and Innovative Therapies, Polytechnic University of Marche.

Identification of T-Complex Protein 1 as a Novel Regulator of the Store-Operated Calcium Channel Orai1 Abundance at the Plasma Membrane

Rawad Hodeify1, Manjula Nandakumar2, Maryam Own2, Raphael J Courjaret2, Satanay Z Hubrack2, and Khaled Machaca2

1Department of Biotechnology, American University of Ras Al Khaimah, UAE
2Department of Physiology & Biophysics, Weill Cornell Medicine, USA

Intracellular calcium (Ca2+) is a key signalling molecule. Store operated calcium entry (SOCE) is a fundamental Ca2+ influx pathway at cell membrane activated in response to intracellular Ca2+ store depletion. The SOCE machinery consists of ER-localized Ca2+ sensor, STIM1, and a Ca2+ selective channel at plasma membrane (PM), Orai1. At steady state, 40% of the total Orai1 protein pool localizes to PM while Orai1 cycles rapidly between PM and intracellular compartments. We discovered that a significant population of intracellular Orai1 pool localizes to a novel sub-PM vesicular compartment. Store depletion results in significant enrichment at PM. To identify Orai1-interacting proteins enriched in sub-PM vesicular compartment, we followed a quantitative proteomic analysis approach. TCP-1 (T -complex protein 1) was identified to be associated with Orai1 in sub-PM vesicular compartment. To functionally assess the role of TCP -1 in Orai1 sub-cellular localization, we knocked down TCP-1 in stable YFP-HA-tagged Orai1 CHO cells using RNA interference. Our data showed significant increase in surface Orai1 upon TCP-1 knockdown. Furthermore, we generated a mutant Orai1 in which the TCP-1-binding homologous region in the intracellular loop was scrambled. This mutant showed decrease binding to TCP-1, and 30% increase in surface Orai1. More importantly, the scrambled Orai1 mutant, when co-expressed with STIM1, showed faster TG-induced puncta formation, increased decay time after ATP-induced SOCE, and faster SOCE-dependent NFAT1 translocation.

In summary, using an unbiased proteomics approach we successfully identified TCP-1 as a novel regulator of Orai1 abundance at PM. Disruption of TCP-1-Orai1 binding resulted in higher plasma membrane residence, faster puncta formation, and faster SOCE development. TCP-1, therefore, plays a key role in Orai1 residence at PM and controlling SOCE and intracellular Ca2+. These results demonstrate that TCP-1 is a novel regulator of Orai1 PM residence and activity, and introduce new directions for design of novel therapeutic strategies targeting SOCE.

Dr. Rawad Hodeify earned his Ph.D. in Interdisciplinary Biomedical Sciences from the University of Arkansas for Medical Sciences, USA, in 2011, and M.S. in Biology from American University of Beirut, Lebanon. He received research training as postdoctoral fellow in Internal Medicine department at University of Arkansas for Medical Sciences and Weill Cornell Medicine before recently joining American University of Ras Al Khaimah as an Assistant Professor of Medical Biotechnology. His research work is focused on regulation of calcium signalling pathways during cellular development and differentiation and dissecting the crosstalk between cycle cell proteins and cell death pathways in kidney injury.

Soluble Receptor for Advanced Glycation End Products (sRAGE) as Potential Biomarker for Diabetic Retinopathy and Chronic Kidney Disease

Umah Rani Kuppusamy1, Wong Foo Nian1, Chua KekHeng1 and Ng Zhi Xiang2

1Department of Biomedical Science, University of Malaya, Malaysia
2School of Biosciences, University of Nottingham, Malaysia

The global prevalence of Type 2 diabetes (T2D) which often leads to dire complications has increased significantly and 17.5% of adult population in Malaysia has T2D. Receptor for advanced glycation end-products (RAGE) is a multi-ligand, cell-surface receptor, which has been widely investigated for its role in the pathogenesis of complications with or without diabetic origin. RAGE also exists as other isoforms which include soluble RAGE (sRAGE) which acts as a decoy receptor that circumvents signal transduction originating from RAGE-ligand binding. Assessment of the levels of sRAGE and oxidative indices in diabetic patients with and/or without diabetic retinopathy (DR) or chronic kidney disease (CKD) and healthy control revealed mixed results. The soluble RAGE level of the patients with DR was significantly higher than healthy controls. Both sRAGE and pentosidine (an oxidative glycation marker) were significantly correlated with diabetes duration. The sRAGE/pentosidine was lower in patients with DR than the healthy controls. Logistic regression analysis revealed positive correlation between sRAGE/pentosidine ratio and the severity of DR and thus, it could serve as a biochemical tool for monitoring the progression of DR. Diabetic patients had significantly higher sRAGE levels in the presence of CKD or DR than in the absence of complications. However, the correlation between sRAGE and renal function in non-diabetic CKD was confounded by other factors but remained highly significant in diabetic CKD even after the adjustment of the potential confounding factors thus, serving as a more convincing indicator of the latter. These findings suggest that sRAGE can serve as a potential biomarker of DR and diabetic CKD. In view of the significant correlations between sRAGE and the severity of these complications, sRAGE could also serve as a tool for monitoring the progression of these complications.

Dr. Umah Rani Kuppusamy is the Head of the Department of Biomedical Science, Faculty of Medicine, University of Malaya. She obtained her PhD in Biochemistry from the National University of Singapore in 1994. Her research interest revolves around free radicals, antioxidants and oxidative stress in diseases with predominant focus on the etiology and complications of diabetes, obesity and mechanism of action of micronutrients on oxidative stress. She has more than 80 publications in ISI-ranked journals, 130 conference papers, medals and awards and several patents to her credit.

Can Phosphoproteomic Analysis Combined to Metabolic and Mutational Phenotyping Help define Individual Profiles of T-ALL Cells?

Sandra Marmiroli1, Laura ANSELMI1, Manuela Zavatti1, Valentina Serafin2, Maddalena Paganin2, Jessika Bertacchini1, Giuseppe Basso2, Benedetta Accordi2 and Xu Huang3

1Department of Biomedical, Metabolical and Neural Sciences, University of Modena and Reggio Emilia, Italy
2Department of Womanʼs and Childʼs Health and Institute of Pediatric Research – Citta della Speranza Foundation, University of Padova, Italy
3Paul OʼGorman Leukaemia Research Centre, College of Medical, Veterinary & Life Sciences, Institute of Cancer Sciences, University of Glasgow, United Kingdom

Prognosis of chemo resistant or relapsed T-ALL patients is still very poor. Several key signaling pathways are deregulated in T-ALL, such as the PI3K/mTOR cascade, downstream of Notch1 mutations (found in >60% patients) or PTEN gene deletion/inactivation. These alterations frequently lead to reprogramming of metabolism, whereby cancer cells display glycolytic features even in normoxic conditions to boost rapid growth and energy demand. In spite of the heterogeneity of this malignancy, however, hitherto most patients are still treated with conventional chemotherapy regimens.

A panel of highly characterized T-ALL cell lines, recapitulating the heterogeneity of T-ALL phenotypes, and primary cells from patients were analyzed by NGS for Notch mutation, by GEP for expression of genes involved in regulation of cell metabolism downstream of the PI3K pathway, and by RPPA for the phospho profile of the PI3K/Akt/mTOR cascade. Analysis of the energy metabolism phenotypes was carried out by the Seahorse XFe96Analyzer. Next, we examined the responses of all cell lines to treatments with drugs blocking PI3K signaling (PF-4691502) and/or glycolysis (2DG).

Overall, our results indicate that cells carrying both Notch1 and PTEN mutations display higher signaling and a more glycolytic phenotype, compared to those with wild type and/or a single mutation. Besides, in these cells 2-DG and PF-4691502 show strong synergistic cytotoxicity and abrogate cell proliferation even at very low concentration, evaluated by CFU assay. Moreover, cells carrying mutant/cleaved Notch1 alone are more sensitive to 2-DG as monotherapy, indicating that Notch1 may be more effective in driving metabolic rewiring. On the other side, cells carrying mutant PTEN alone are highly sensitive to PF-4691502, indicating a prevailing role of the signaling over activation in these cells.

These results, though preliminary, suggest that mutational and phosphorylome analysis of T-ALL correlated to metabolic phenotypes can allow to define individual profiles and to predict specific treatments effectiveness.

Dr. Sandra Marmiroli has done her PhD in Cellular and Molecular Biology and is an Associate Professor of Histology. She is the author or co-author of 71 full-length peer-reviewed papers (Scopus), 1570 citations (Scopus), H-index 27 (Scopus + WOS). Her main research topics include “Identification of glycolytic vs. oxidative cellular phenotypes and their modulation by the PI3K pathway in primary blast cells from leukemia patients”. “Definition of the phosphorylome of primary blast cells from leukemia patients, and its modulation by the PI3K pathway:” “Targeted therapy in hematological malignancies” (Bertacchini et al., Leukemia 2014; Serafin et al., Leukemia 2017)

Proliferation of Porcine Adipose Derived Stem Cells in Herbal Scaffolds

Franca Nneka Alaribe1, Shirley CKM Motaung1 and Cornelius C Ssemakalu2

1Department of Biomedical Sciences, Tshwane University of Technology, South Africa
2Department of Biotechnology, Vaal University of Technology, South Africa

The significance and the potentials of in vitro cell culture studies are great considering the need for more cost efficient development of new drugs, time efficient treatment of cancer patients, and an understanding of developmental biology and mechanisms of stem cell differentiation. Cells, growth factors and scaffolds are the fundamental issues for tissue engineering. If porcine derived adipose stem cells (pADSCs) can effectively proliferate and differentiate when cultured on herbal scaffolds, this makes it a potential candidate for in vivo environment with possible profound impact on therapeutic application of herbal scaffolds. This study evaluated the in vitro differentiation capacity and anti-inflammatory effect of fabricated herbal scaffolds on pADSCs. To this effect, herbal scaffolds were developed by incorporating medicinal plant extracts (Eucomisautumnalis and Pterocarpusangolensis) and natural biopolymers (Alginate and chitosan) using lyophilisation technique. A standard sterility test on the scaffolds before in vitro use showed the ultraviolet radiation with 75% (v/v) ethanol to be suitable. pADSCs cultured on the herbal scaffolds were further monitored for in vitro proliferation and differentiation using different biological, immunological and genetic techniques. The identity of pADSCs were confirmed by positive FACs analysis of mesenchymal stem cell surface markers CD44, CD90 and CD105 (≥ 85%). Their multi potency was further evaluated by trilineage differentiation of pADSCs toward adipocyte, osteocyte and chondrocyte with histology staining. Scanning electron microscope (SEM) revealed that the herbal scaffolds possess an extremely porous structure than control (non-herbal scaffold). SEM and immunofluorescence results also revealed more attachment of ells at day 7, 14 and 21 on herbal scaffolds than non-herbal scaffolds. ELISA assay with IL-6 and gene expression of collagen type 11, alkaline phosphate, osteocalcin and osteopontin with RT-PCR confirmed the anti-inflammatory nature and mRNA expression/chondrogenic nature of the pADSCs cultured with the herbal Scaffolds media. This study indicates that pADSCs would have great therapeutic potential as seeding cells for in vivo transplantation to treat various inflammatory diseases and bone injuries when co-applied with medicinal plants and biopolymers.

Dr. Franca Nneka Alaribe, PhD Biomedical Sciences, University of Ferrara Italy, specializing in cellular/molecular pathology in cancer and degenerative diseases; MSc Molecular Biology/Biotechnology, Vrije University Brussels Belgium; BSc(Ed) Biology, University of Nigeria Nsukka. She is a researcher for over 10 years with high competence in biomedical research and has coordinated many research projects for NRF, THRIP, Bone South Africa and OSTA involving biomaterials. Presently she is working on stem cells and scaffold fabrication with South African medicinal plants. She has published more than 12 research works and academic papers in international peer reviewed journals, founder/editorial manager of Journal of Advances in Biomedical Studies. Editor of several peer reviewed journals and fellow of many scientific organisations.

Prognostic Value of the Expression of Endogenous Hypoxia Associated Proteins Hypoxia Inducible Factor-1 Alpha (HIF-1α) and Carbonic Anhydrase Isoform 9 (CAIX) Expressions in Breast Carcinoma

Ola A Harb1, Loay M Gertallah1 and BasantSh El Shafaay1

1Department of Pathology, Zagazig University, Egypt
2General Surgery, Zagazig University, Egypt
3Department of Clinical oncology & nuclear medicine, Zagazig University, Egypt

Background: Hypoxia has been found to be related to malignant initiation, progression, increasing the occurrence of metastasis and therapy resistance in many cancer types, which made a real need for discovering drugs that could antagonize the bad effect of hypoxia in cancer, decide which patients will have benefit from such anti-hypoxia therapy then to monitor response to therapy, especially in breast carcinoma. It is important to detect degree of hypoxia in each cancer that could be done by evaluation of the expression of hypoxia-associated protein in cancer biopsies e.g. hypoxia inducible factor-1 alpha (HIF-1α) and carbonic anhydrase IX (CAIX) and their detailed role in breast cancer is still uncertain and gives conflicting results.

Aim of the Work: Was to evaluate HIF-1α and CAIX expressions in breast carcinoma, correlating their expressions with each other, with presence of lymph node & distant metastases, with recurrence free and overall survival rates of breast cancer patients.

Methods: We evaluated HIF-1α & CAIX expressions in sections from 90 paraffin blocks of breast carcinoma using immunohistochemistry. We analyzed correlations between their levels of expressions, clinic-pathological and prognostic parameters of our patients.

Results: HIF-1α and CAIX positive expression in breast carcinoma was related to advanced stage, presence of lymph node metastases, HER2 amplified and triple negative molecular subtypes (p<0.001), higher tumor grade (p= 0.001 & 0.02 respectively) and negative ER (p= 0.005 & 0.008 respectively) & PR (p= 0.009 & 0.027 respectively) hormonal receptors, The expression of both markers was significantly positively correlated with each other (p<0.001). HIF-1α and CAIX positive expression in breast carcinoma was associated with shortened recurrence free and overall survival rates (p<0.001).

Conclusion: HIF-1α and CAIX are markers of poor prognosis of breast carcinoma patients.

Keywords: Breast carcinoma, hypoxia; HIF-1α; CAIX; immunohistochemistry; prognosis

Dr. Ola A Harb, MD; completed her Pregraduate Medical Education (December 2005) in M.B.B.CH., with Total grade-Excellent from Zagazig University, Egypt. She obtained her Postgraduate/M.Sc (May, 2010) & M. D. (January 2015) in pathology from Zagazig University, Egypt. Dr. Ola is presently working as a Lecturer, at Department of pathology, Faculty of Medicine, Zagazig University, Egypt.

Papillary Thyroid Carcinoma

Idania Teresa Mora López

Enrique Cabrera General Teaching Hospital, Cuba

The papillary thyroid carcinoma is the most frequent endocrine neoplasia and the one with the highest presentation in thyroid carcinomas, prevalence (5-6.5%), predilection for the female sex, second to third decades of life and increase in aging, influences predisposing factors such as head and neck radiation, thyroid nodule background, representing (5-15%), family history and the association with Multiple Endocrine Neoplasia type 2, Cowden syndromes, Gardner, Garney complex or unknown etiology.

Diagnosis based on questioning, specifying exposed history, physical examination, determining consistency, location of the nodule with respect to the gland, size, regional lymph nodes, supraclavicular, growth pattern and complementary studies such as ultrasound that expresses whether it is solid, hypoechoic, irregular borders, microlobulated, microcalcifications, absence of safety halo, thick and incomplete halo, determination of thyroid function with Thyroid Stimulating Hormone (TSH), Thyroglulin and Fine Needle Aspiration Cytology, which according to the Bethesda System confirms carcinoma (97-99%) and diagnosis definitive with freezing biopsy and adenomegalies if they were removed.

Treatment based on total thyroidectomy, later treatment with thyroid hormones (suppressive dose), follow-up with (TSH) per month, one year Gammagraphy and thyrogluline, according to the annual follow-up results in the first 5 years and every 2 years in the following 5 years, every 5 years the rest of the life, if it detects an area in the neck, perform treatment with 131IODO assess every 6 months and repeat at the same time until there is negativity and always doing thyroglycine. Treatments such as chemotherapy and tyrosine kinase inhibitors in non-operable and advanced cases.

Dr. Idania Teresa Mora López Graduated in Medicine in 1990, Faculty of Medicine Julio Trigo Lopez, belonging to the Medical Sciences University of Havana. She obtained her First Degree Specialist in Comprehensive General Medicine and Specialist of First and Second Degrees in Endocrinology. And Diploma in Higher Medical Education, Master of Science Satisfactory Longevity & as a Aggregate Investigator. Presently she is Adjunct Assistant Professor of the Dr. Enrique Cabrera Medical Sciences University and the Latin American School of Medicine. She holds wide publications of 15 national / international.

Developing Patient-Specific Direct Neuronal Reprogramming for Modeling Neurodegenerative Disorders

J Drouin-Ouellet1, S Lau1, PL Brattås1, DR Ottosson1, K Pircs1, D Grassi1, J Jakobsson1, M Parmar1, LM Collins2, R Vuono2, A Andersson Sjoland3, G Westergren-Torsson3, C Graff4,5, L Minthon6, H Toresson6 and RA Barker1,2

1Department of Experimental Medical Science, Wallenberg Neuroscience Center, Division of Neurobiology and Lund Stem Cell Center, Lund University, Sweden
2John van Geest Centre for Brain Repair & Department of Neurology, Department of Clinical Neurosciences, University of Cambridge, UK
3Department of Experimental Medical Science, Lund University, Sweden
4Karolinska Institute, Department of NVS, Center for Alzheimer Research, Division for Neurogeriatrics, Sweden
5Karolinska University Hospital, Dept of Geriatric Medicine, Sweden
6Clinical Memory Research Unit, Department of Clinical Sciences Malmo, Lund University, Sweden

Direct conversion of adult human fibroblasts into mature and functional neurons, termed induced neurons (iNs) was achieved for the first time five years ago. This technology offers a shortcut for obtaining patient and disease specific neurons for disease modeling, drug screening and other biomedical applications. Despite their great promise, reprogramming roadblocks have prevented the generation of iNs at a sufficiently high yield from adult dermal fibroblasts, which has significantly limited the adoption of this technology. To overcome this, we have developed a new highly efficient dual promoter-based vector system that results in efficient co-delivery of the two reprogramming factors Brn2a and Ascl1 in combination with either neuron specific microRNAs or the inhibition of the RE1-silencing transcription factor (REST). Global gene expression analysis showed that while both strategies resulted in induction of a neuronal program and similar level of neural conversion, the inhibition of REST induces the expression of additional genes that are related to neuronal identity and function. Based in this, we developed an optimized one-step method to efficiently reprogram dermal fibroblasts from elderly individuals using a single vector system and demonstrate that it is possible to obtain iNs of high yield and purity from aged individuals, including Parkinsonʼs disease patients. We are now evaluating the conversion capability of these skin fibroblasts and our preliminary results suggest that iN cells from sporadic PD patients reprogram in a similar fashion as that of healthy individuals and could thus serve as a tool to model intracellular pathological features associated with PD.

Dr. Drouin-Ouellet has obtained her Ph.D. in Neurobiology at Laval University (Quebec City, Canada) in 2012. She is currently working at Lund University (Lund, Sweden) where she has developed a simple and efficient method for direct neuronal conversion of adult human dermal fibroblasts from patients with neurodegenerative diseases. She is now conducting research aiming at further advancing the direct neuronal reprogramming technology to provide patient specific based systems that faithfully recapitulate disease-associated phenotypes of neurodegenerative disorders in vitro.

Treatment of Diabetes and Obesity with CRISPR-Mediated Genome Editing in Epidermal Progenitor Cells

Xiaoyang Wu1, Jiping Yue1, Xuewen Gou1, Cynthia Li1 and Barton Wicksteed2

1Ben May Department for Cancer Research, University of Chicago, USA
2Division of Endocrinology, Diabetes and Metabolism, University of Illinois, USA

Somatic gene therapy with current genome editing technology provides a promising therapeutic approach for treatment of a variety of otherwise terminal or severely disabling diseases. The human skin is a tempting target for genetic engineering as it is the biggest and most accessible organ in our body. Moreover, skin epidermal stem/progenitor cells are easy to obtain and expand in vitro, and extraordinary advances have been made in the development of epidermal autograft or tissue-engineered skin equivalents for permanent skin regeneration in clinics. In this report, by combining CRISPR-mediated genome editing with epidermal progenitor cell platform, we develop skin graft with controllable release of GLP1 (glucagon-like peptide-1), a critical incretin that regulates blood glucose homeostasis, and demonstrate its therapeutic effect in vivo by reducing glycemic excursions in diet-induced obese and diabetic mice. Taken together, our study lays the essential groundwork for development of long-lasting and safe gene therapy approach for combating obesity and diabetes, and unravels the clinical potential for genome editing in skin epidermal progenitor cells.

Dr. Xiaoyang Wu received his PhD in 2000-2006 from Cornell University. He was post doctorate fellow from Rockefeller University, in 2006-2011. From 2011-present, is working as an Assistant Professor at University of Chicago, USA.

Stem Cells Improve Kidney Function and Remodelling in CRS Type II

Chiara Castellani1, Marny Fedrigo1, Andrea Di Francesco1, Marialuisa Valente1, Gaetano Thiene1, Annalisa Angelini1, Regina Tavano2, Emanuele Papini2, Michela Pozzobon3 and Giorgio Vescovo4

1Departmentof Cardiac, Thoracic and Vascular Sciences, University of Padua, Italy
2Department of Biomedical Sciences, University of Padua, Italy
3Department of Women and Children Health, University of Padua, Italy
4Internal Medicine, S. Antonio Hospital, University of Padua, Italy

Background: We investigate the effects of human amniotic fluid stem cells (hAFS) and rat adipose tissue stromal vascular fraction GFP-positive cell (rSVC-GFP) therapy in a monocrotaliner at model of cardio-renal syndrome type II (CRSII).

Methods: RHF was induced by monocrotaline (MCT) in Sprague-Dawley rats. Three weeks later, four million of hAFS or rSVC-GFP were injected via tail vein. BNP, sCreatinine, kidney and heart NGAL and MMP9, sCytokines, kidney and heart apoptosis (TUNEL technique) were studied. Stem Cells (SC) engraftment was detected with immunofluorescence.

Results: SC treated rats showed a significant reduction of serum NGAL and Creatinine (NGAL 335.6±92.60 sCrea 0.36±0.05, p= 0.01) compared to CHF rats. In both hAFS and rSVC-GFP group, kidney protein expression of NGAL was significantly lower than in CHF group (SC 2.6*106±1.2*106 vs HF 5.1*106 ± 1.5*106 A.U., p=0.0008) and similar to that to controls. In both hAFS and rSVC-GFP treated rats, we observed a substantial number of SC engrafted in the medulla and differentiated in tubular cells. Apoptosis was significantly decreased (hAFS 10.29±10.81 and rSVC-GFP 24.82±25.19 cells/mm2, p=0.05 vs CHF) and similar to controls (9.85±7.2 cell/mm2). TUNEL-positive cells were mainly located in the kidney medulla. Pro-inflammatory cytokines were down regulated in SC-treated groups (p=0.05 vs CHF) and similar to controls. In SC treated rats, kidney and heart tissue NGAL was not complexed with MMP9 as showed in CHF groups, suggesting inhibition of MMPs activity.

Conclusion: SC treatment produced improvement in kidney function in rats with CRSII. This may be the results of tubular regeneration due to SC engraftment, decrease tubular cells apoptosis and mitigation of pro-inflammatory milieu. Reduction of NGLA-MMP) complexion mainly to decrease MMPs activity with prevention of further negative heart remodeling

Dr. Chiara Castellani, is the Assistant Professor in Technical Sciences of Laboratory Medicine. (University teaching of “Cyto and histopathological diagnosis” and part of courses of Pathological Anatomy.) Dr. Chiara got her Ph.D. in Cardiovascular Pathology (2007) & achieved thanks to a research on “Cardiac and non-cardiac stem cells in heart transplant and cardiac hypertrophy remodelling”. She completed her Specialty training in Clinical Biochemistry (March 2013). She has been a visiting research fellow at the Washington University in Seattle, WA (2006), where she worked under the supervisor ship of Prof. Charles Murry on the heart remodeling and stem cells.
Research lines: i) Stem cells and cardiac remodelling; ii) stem cells and cardio renal syndrome; iii) cardiac amyloidosis; iv) Cardiac allograft vasculopathy; v) Exosomes and miRNA in heart and kidney transplantation.
The outcome of her research was presented through oral presentations and posters during many national and international meetings. She had published papers in international ISI journals and one book chapter (HI=11 and citations)

Treating Diet-Induced Obesity with Adipose-Derived Mesenchymal Stem Cells: Preclinical Evidence-Based Approach

Fatima Saleh1, Leila Itani2, Marwan El Ghoch2,3, Simona Calugi3, and Riccardo Dalle Grave3

1Department of Medical Laboratory Sciences, Beirut Arab University, Lebanon
2Department of Nutrition and Dietetics, Beirut Arab University, Lebanon
3Department of Eating and Weight Disorders, Villa Garda Hospital, Italy

Introduction: Obesity has become a major healthcare issue over the past two decades, but its treatment is not yet optimal, especially in the long-term. The current status pressed the need for a novel treatment for obesity and its associated comorbidities. Stem cell-based therapy is emerging as a promising therapy for diet-induced obesity. The purpose of this study is to highlight the efficacy and safety of adipose derived stem cells (AD-MSCs) on obesity and related comorbidities in animal models, in order to establish the feasibility of translation into the clinical setting for a possible treatment in humans.

Methodology: A Systematic review was conducted in adherence to the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guidelines, and the protocol was registered in the PROSPERO registry (CRD42017077347).

Findings: The initial search retrieved 578 papers, and seven articles met the inclusion criteria. Strong evidence reported the positive effect of AD-MSCs on obesity treatment in terms of body weight, glucose metabolism homeostasis, lipid profiles, non-alcoholic fatty liver disease and systemic inflammation.

Conclusion: This study demonstrates the promising beneficial effects of AD-MSCs on obesity and obesity-related diseases such as type 2 Diabetes and dyslipidemia in animal models. However, more studies should be performed to understand their mechanism of action and to overcome some methodological limitations evidenced in our systematic review before moving forward to consider AD-MSCs transplantation into human.

Dr. Fatima Saleh is an Assistant Professor in the Faculty of Health Sciences at Beirut Arab University (BAU) since 2013. She obtained her BSc in Pharmacy in 2004 from BAU and MSc in Pharmacology and Biotechnology from UK. Dr. Saleh received her PhD degree in Biomedical Sciences from University of York, UK. Her current research interests focus on Mesenchymal stem cell research as well as identification of natural compounds with antidiabetic, anti-oxidant or antimicrobial activity. She published in peer-reviewed journals and presented her work in many international conferences.

Primary Cancer Cell Cultures for Personalized Cancer Therapy

Secil Yilmaz

Genome and Stem Cell Center, Erciyes University, Turkey

Even though the last ten years have seen major discoveries in cancer research, cancer is still a global health issue. Most of cancer research are based on cancer cell lines and animal models. Immortal cancer cell lines are the most widely used models for investigating cancer biology but cell lines poorly represent the tumor heterogeneity of the cancer patients. The average rate of successful translation from animal models to clinical cancer trials are very low as a result of the traditional animal cancer models are limited in their ability to recapitulate the progression of cancer in human. Being able to transfer results from the lab to clinical studies and beyond is crucial. Primary cancer cells, by their very nature, are a useful tool in developing novel cellular therapies for personalized medicine and autologous treatments, by reason of the human primary cancer cells derived individual patients can closely mimic the origin of the disease. Each cancer patient needs to be considered as an independent individual with a unique disease since even in a same cancer disease, every single cancer patient has different genotypical/ phenotypical features. Therefore, the best personalized therapy should be the main subject of the cancer research. In addition, the application of primary cancer cell culture models has the potential to develop the three-dimensional (3D) tumor models for future cancer therapies and to determine biomarkers for diagnosis and treatments.

Dr. Yilmaz is Assistant Professor in Genome Stem Cell Center at Erciyes University. She worked with nobel laureate 2015, Aziz Sancar, at University of North Carolina during the PhD studies. Her current research involves cellular-based approaches to personalize the cancer theraphy.

Development of a 3D Cell Culture Model Based on Biocompatible Polymeric Scaffolds Engineered with Human Mesenchymal Stromal Cells (Mscs) for Skin, Cartilage and Bone Regenerative Therapy

Re Federica1*, Russo Domenico1, Perucca Simone1, Sartore Luciana2, Bignotti Fabio2, Almici Camillo3, Rodella Luigi4, Salmeron-Sanchez Manuel5, Sardini Emilio6, Guizzi Pierangelo7, Calzavara Pinton Piergiacomo8, Paganelli Corrado9 and Magrini Stefano Maria10

1Unit of Blood Diseases and Adult Bone Marrow Transplantation - Department of Clinical and Experimental Sciences - University of Brescia, Italy
2Materials Science and Technology - Department of Mechanical and Industrial Engineering - University of Brescia, Italy
3Laboratory of Manipulation and Cryopreservation of Stem Cells - Immunohaematology and Transfusion Medicine Service - ASST Spedali Civili di Brescia, Italy
4Section of Anatomy - Department of Clinical and Experimental Sciences, University of Brescia, Italy
5Chair of Biomedical Engineering - University of Glasgow, UK
6Electrical and Electronic Measurements - Department of Information Engineering - University of Brescia, Italy
7OU Orthopedics and Traumatology - ASST Spedali Civili di Brescia (Presidio of Gardone Val Trompia), Italy
8Division of Dermatology - Department of Clinical and Experimental Sciences - University of Brescia, Italy
9Odontostomatological Diseases - Department of Medical-Surgical Specialties, Radiological Sciences and Public Health - University of Brescia, Italy
10Diagnostic Imaging and Radiotherapy - Department of Medical-Surgical Specialties, Radiological Sciences and Public Health - University of Brescia, Italy

Regenerative medicine aims to restore normal function by repairing or replacing damaged cells and tissues in patients. The efficacy and the success of regenerative medicine depends on many factors including the manner by which the cells are organized in the new tissue that must be able to mimic the structure and function of the original one. Current treatments for articular cartilage damage, bone defects and skin lesions are quite challenging and they show limited repair and regeneration following injury.

This project aims at developing an innovative 3D cell culture model composed of Mesenchymal Stromal Cells (MSCs) and biocompatible, safe, resorbable, polymeric scaffolds for studying the mechanisms involved in tissue repair and treatment of chronic skin conditions, cartilage defects and bone lesions.

For this purpose, hydrogel-forming polymers scaffolds of natural origin were firstly characterized for their mechanical and morphological properties, stability and degradability. Secondly, MSCs were seeded on these scaffolds and optimal culture conditions for MSCs expansion were defined. Preliminary results suggest that hydrogel scaffolds seem to be an optimal model to host MSCs.

Currently, we are exploring the effects of scaffolds on modulating MSCs behaviour, in particular in the matter of how scaffolds can affect MSCs proliferation and differentiation. Moreover, efforts are being made to implement this 3D culture system with autonomous sensors in order to have an objective way to monitor tissue restoration.

This project will provide new useful insights to define novel innovative therapeutic approaches, thus making this research translatable within the clinical scenario.

Dr. Federica Re has obtained her Degree in Medical Biotechnology from 2010 to 2015 at the Department of Molecular and Translational Medicine of the University of Brescia, Italy. She is doing a PhD in Technology for Health at the department of Information Engineering at the University of Brescia, Italy. She spent several months at the Department of Biomedical Engineering at the University of Glasgow, Scotland during her PhD. Her main research interest is focused on the study of the human Mesenchymal Stem Cells applicationsin combination with biocompatible and bioresorbable scaffolds for a cost-effective regenerative therapy translatable into the clinical practice.

A New Polylipoic Acid-Based Nano-Platform for Heart Diseases Treatment

Chiara Castellani1, Marny Fedrigo1, Annalisa Angelini1, Regina Tavano2, Emanuele Papini2, Fabrizio Mancin3 and Jakubwaldemar Trzcinski3

1Department of Cardiac, Thoracic and Vascular Sciences, University of Padua, Italy
2Department of Biomedical Sciences, University of Padua, Italy
3Department of Chemical Sciences, University of Padua, Italy

Nanomedicine, that is the application of nanotechnologies to medicine, attracts an enormous interest. The aim of this study is to evaluate the safety, feasibility and biocompatibility of a new polylipoic acid-based nano-platform (NPs) for heart diseases treatment.

Material and Methods: The lipoic acid polymerization reaction developed by Matile et al. has been used to create highly cross-linked polymeric nanoparticles. Dimeric and trimeric derivatives of lipoic acid with different spacers have been synthesized and used to produce surfactant stabilized nanoemulsions. Thiol-initiated polymerization of the microemulsions have been lead to the formation of a highly reticulated polymeric nanostructure. Loading of active molecules (such as rhodamine) has been obtained either by entrapment or by copolymerization of appropriate lipoic acid derivatives in the nanoparticle matrix. HeLa cells, Raw 264.7 cells, purified human leukocytes and human monocyte-derived. Macrophages were incubated with different concentrations of nanoparticles (up to 200 µg/ml) for 24h hours, the vitality was assessed by MTS assay and the association of NPs to the cells was assessed by cytofluorimetry. Red blood cells, purified by buffy coats of healthy donors, were incubated for 24h with different concentrations of nanoparticles (up to 200 µg/ml) and then the possible lysis of the red blood cells has been monitored by the release of hemoglobin measured at 540 nm.

18 male health Sprague-Dawley rats were injected with 10mg/Kg of NPs via vein tail. 7 male health rats were taken as controls. Rats were sacrificed 1hour, 3 hour, 24 hours 3 and 7 days after NPs injection and blood and organs were collected. Section of 2mm of lung, heart, liver, kidney and spleen were analyzed by Alliance 2.7 3D software to identify and quantify NPs-rhodamine conjugated. NPs localization in tissues was identify by confocal microscopy.

Results: We found that in vitro this new polylipoic acid-based nano-platform didnʼt exert any toxicity towards all the different cell types we used, even at high concentrations (200 µg/ml); NPs associate at low levels to all the tested cells, but they are captured by human macrophages at high levels. Moreover, NPs didnʼt induce red blood cell lysis. In vivo, NPs fluorescence was identify immediately after injection in the heart. Moreover the heart was able to retain the NPs until 7 days after injection. The heart demonstrated to have a low clearance of NPs. Liver and kidney showed NPs fluorescence clearance at 3h after injection. Confocal microscopy showed that NPs are localize in the interstitium of organs and in the endothelial cells. NPs did not show any toxicity in the rats.

Conclusion: Heart is able to retain NPs injected in blood flow for up to 7 days without evident negative side effects. These preliminary data suggest that this new nano-platform formulation could be used to target heart diseases and for therapeutic drug delivery.

Dr. Chiara Castellani, is Assistant Professor in Technical Sciences of Laboratory Medicine. (University teaching of “Cyto and histopathological diagnosis”and part of courses of Pathological Anatomy.) Dr. Chiara got her Ph.D. in Cardiovascular Pathology (2007) & achieved thanks to a research on “Cardiac and non-cardiac stem cells in heart transplant and cardiac hypertrophy remodelling”. She completed her Specialty training in Clinical Biochemistry (March 2013). She has been a visiting research fellow at the Washington University in Seattle, WA (2006), where she worked under the supervisor ship of Prof. Charles Murry on the heart remodeling and stem cells.
Research lines: i) Stem cells and cardiac remodelling; ii) stem cells and cardio renal syndrome; iii) cardiac amyloidosis; iv) Cardiac allograft vasculopathy; v) Exosomes and miRNA in heart and kidney transplantation.
The outcome of her research was presented through oral presentations and posters during many national and international meetings. She had published papers in international ISI journals and one book chapter (HI=11 and citations)

Expression of Cox-2, Cyclin D1 and P21 in Colorectal Cancer Patients and their Clinicopathological and Prognostic Significances

Ola A Harb1, Mariem A Elfeky1 and Loay M Gertallah2

1Department of Pathology, Zagazig University, Egypt
2General Surgery, Zagazig University, Egypt

Background: Cyclooxygenase-2 (COX-2), play an important role in inflammation, carcinogenesis and cell cycle alterations. It is incriminated in cancer progression by causing dysregulation of normal cell cycle control. Cyclin D1 plays a vital role in cancer cell cycle progression. The activity of cyclin D1 can be blocked by CDK inhibitors, including p21 (cyclin-dependent kinase inhibitor-1A, CDKN1A, CIP1) that plays a role in regulating cell cycle.

The purpose of this study: Was as to evaluate expression of COX-2, Cyclin D1 and P21 in colorectal cancer patients, analyze the relationship between their expression, clinicopathological criteria and the prognosis of patients.

Methods: Expressions of Cox-2, Cyclin D1 and P21 were evaluated in 60 paraffin blocks colorectal cancer patients that were followed up for 3 years. The relationship between their level of expressions and prognosis of patients was analyzed.

Results: Cox-2 & Cyclin D high expression was positively correlated with higher grade, advanced stage, presence of lymph node & distant metastasis and Duke stage (P=0.000).

P21 high expression was negatively correlated with presence of lymph node metastases, higher grade (p=0.002), advanced stage, presence of distant metastasis and advanced Duke stage (P=0.001). We found a direct relationship between Cox-2 and Cyclin D 1, an inverse relationship between Cox-2 and P21, and an inverse relationship between Cyclin D 1 and P21 (P<0.001).

Cox-2 & Cyclin D1 over expression and P21 low expression were positively associated with higher incidence of tumor recurrence (P=0.04, 0.000 respectively), higher incidence of cancer specific death (p=0.002, 0.004 & 0.000 respectively) but no significant correlation with response to therapy with all markers.

Conclusion: High levels of expression of Cox-2 & Cyclin D1are markers of poor prognosis, while high level of expression of P21 is a marker of good prognosis in colon cancer patients these results suggest that loss of control of cell cycle check points is a common occurrence in CC, and regulation in cell growth control and tumor suppression.

Keywords: Cox-2, Cyclin D1, P21, colon cancer patients, immunohistochemistry, prognosis

Dr. Ola A Harb, MD; completed her Pregraduate Medical Education (December 2005) in M.B.B.CH., with Total grade-Excellent from Zagazig University, Egypt. She obtained her Postgraduate/M.Sc (May, 2010) & M. D. (January 2015) in pathology from Zagazig University, Egypt. Dr. Ola is presently working as a Lecturer, at Department of pathology, Faculty of Medicine, Zagazig University, Egypt.

Frontiers of Induced Pluripotent Stem Cells: From Bench to Bedside

Rosy Joshi-Mukherjee

Center for Integrative Research on Cardiovascular Aging (CIRCA), Aurora Research Institute, USA

Stem cell biology has gained tremendous interest in regenerative medicine, offering the hope of new therapies for otherwise intractable disease. Regenerative medicine involves the repair and regeneration of tissues for therapeutic purposes, such as replacing bone marrow in leukemia, cartilage in osteoarthritis or cells of the heart after a heart attack. Today advances in basic and clinical research make tissue regeneration feasible. This course is tailored for researchers with an interest in learning about current biological induced pluripotent stem research in a highly interactive setting. It aims to equip early researchers with the fundamental and cutting edge knowledge and critical understanding necessary for research in stems cells and the implications of induced pluripotent stem cell (iPSC) research for medicine and society. Participants will learn through multiple interfaces such as lectures, discussions, small group activities, and feedback from other participants. Explain the advantages and disadvantages of iPSCs as compared to embryonic stem cells. Describe various methods for creating iPSCs, including transfection methods using plasmids, retroviruses, small molecules, and adenoviruses. Discuss the advantages and disadvantages of each. We will explore basic mechanisms of how iPSC differentiate into specific tissues in response to a variety of biologic signaling molecules. We will discuss the use of such factors for in vitro tissue production. For example, bone morphogenetic proteins can be used in vitro to drive the differentiation of adult stem cells towards bone and heart. Discuss various in-vitro approaches for production of beating heart cells. We will also discuss how studies of the developmental, cellular and molecular biology of regeneration have led to the discovery of new drugswith examples from cutting-edge research. Discuss the clinical uses of iPS cells. Describe issues that must be resolved before iPS cells can safely be used in human cell-based therapy.

Dr. Joshi-Mukherjee developed “Heart-on-µGMEA-Chip Model for cardiomyopathy” using cutting-edge technology namely, induced pluripotent stem cells and multielectrode array. The model reliably predicts cardiac cell death or altered electrophysiology phenotypes such as prolongation of action potential duration and early after depolarizations (EADs). She presented the research at Gordon Research Conference on Cardiac Arrhythmia and Mechanisms at Ventura, CA, United States, in 2017. During postdoctoral fellowship at Johns Hopkins University Dr. Joshi-Mukherjee received T32 fellowship for developing human induced pluripotent stem cell (hiPSC) lines from patient harboring sodium channel mutations associated with long QT 3 syndrome to investigate the mechanisms contributing to arrhythmia.